Transfection Reagents |
Featured Reagents
i-Fect ™ -A novel cationic lipid formulation specifically designed for efficient delivery of 27mer DsiRNAs(dicer substrate small Interfering RNAs)& 21mer siRNAs (small interfering RNAs) in vitro and in vivo.
n-Fect™ -A cationic lipid that has been specifically formulated for nervous system applications. n-Fect provides higher transfection efficiency than other commercially available broad-spectrum transfection reagents for glial cells, neuronal cell lines, and certain primary neuronal cultures.
n-Blast™ -A broad-spectrum transfection reagent successfully used in many cell types commonly used by neuroscientist.
pn-Fect™ -The latest advance in transfection technology for primary neuronal cells. This unique reagent provides ultra-high plasmid DNA delivery efficiencies and low cytotoxicity compared to competitive reagents.
Related Reagents:
p-Fect™ -Designed to delivery plasmids, DNA or RNA to hard to transfect Cell Lines.
pro-Fect™ -Is a unique lipid-based formulation that allows the delivery of proteins, peptides or other bioactive molecules into a broad range of cell types.
Penatratin-1™ -A peptide for delivering small molecules into Neurons and other cells. MP Biomedical is the manufacturer of Penetratin-1
***Delivery of siRNA in vivo & in vitro***
Philippe Sarret , Louis Doré-Savard1, Nicolas Beaudet. Direct Application of siRNA for In Vivo Pain Research. From: RNA Interference: From Biology to Clinical Applications. Series: Methods in Molecular Biology. Volume: 623; Pub. Date: May-01-2010; Page Range: 383-395; DOI: 10.1007/978-1-60761-588-0_25.
...iFect transfection reagent solution (Neuromics, Minneapolis, MN). .... The selected transfection agent is the cationic lipid i-Fect. ...
Pascal Fossat, Eric Dobremez, Rabia Bouali-Benazzouz, Alexandre Favereaux, Sandrine S. Bertrand, Kalle Kilk, Claire Léger, Jean-René Cazalets, Ülo Langel, Marc Landry, and Frédéric Nagy. Knockdown of L Calcium Channel Subtypes: Differential Effects in Neuropathic Pain. The Journal of Neuroscience, January 20, 2010, 30(3):1073-1085; doi:10.1523/JNEUROSCI.3145-09.2010
...the siRNAs (2 µg) were solubilized in 10 µl of i-Fect reagent (Neuromics) following Neuromics instructions and published protocol (Luo et al., 2005)...
Yan Chen, Cheng Yang, and Zaijie Jim Wang. Ca2+/Calmodulin-Dependent Protein Kinase II Is Required for the Initiation and Maintenance of Opioid-Induced Hyperalgesia. The Journal of Neuroscience, January 6, 2010, 30(1):38-46; doi:10.1523/JNEUROSCI.4346-09.2010.
...KN93 and KN92 were administered intrathecally by percutaneous puncture through the L5-L6 intervertebral space, as described previously (Hylden and Wilcox, 1980; Chen et al., 2009). A lateral tail flick was considered as success of the intrathecal injection. To inhibit CaMKII, CaMKII was targeted by small interfering RNA (siRNA). Four days after morphine pellet implantation, mice were treated with CaMKII siRNA (5'-CACCACCAUUGAGGACGAAdTdT-3', 3'-dTdTGUGGUGGUAACUCCUGCUU-5') (Zayzafoon et al., 2005) or Stealth RNAi negative control (Invitrogen) (2 µg, i.t., twice per day for 3 consecutive days). These oligos were mixed with the transfection reagent i-Fect (Neuromics), in a ratio of 1:5 (w/v) (Luo et al., 2005). Mechanical and thermal sensitivity tests were performed daily...
Joseph George, Naren L. Banik, Swapan K. Ray. Combination of hTERT Knockdown and IFN-γ Treatment Inhibited Angiogenesis and Tumor Progression in Glioblastoma. Clin Cancer Res 2009;15(23):7186–95
...with i-Fect transfection reagent (Neuromics) to obtain 5 μg DNA/10 μL of injection volume...
Christian Ndong, Amynah Pradhan, Carole Puma, Jean-Pierre Morello, Cyrla Hoffert, Thierry Groblewski , Dajan O’Donnell, Jennifer M.A. Laird. Role of rat sensory neuron-specific receptor (rSNSR1) in inflammatory pain: Contribution of TRPV1 to SNSR signaling in the pain pathway. PAIN 143 (2009) 130–137.
...For experiments in which siRNA was delivered by bolus injections, 10 ul of siRNA or vehicle was injected directly into the intrathecal catheter once daily for 4 days. In this case, siRNAs were prepared immediately prior to administration by mixing the RNA solution (200 uM in annealing buffer) with the transfection reagent i-FectTM (Neuromics) at a ratio of 1:4 (w:v) for a final siRNA/ lipid complex concentration of 2 ug/10 ul...
Suneeta Tumati, Tally Largent Milnes, Henry I. Yamamura, Todd W. Vanderah, William R. Roeske and Eva V. Varga. Intrathecal Raf-1-selective siRNA attenuates sustained morphine-mediated thermal hyperalgesia. doi:10.1016/j.ejphar.2008.10.033
...siRNAs stock solutions (100 μM) were prepared in double distilled RNAse free water and stored in aliquots at −80 °C. For intrathecal treatment, aliquots of the stock solution (2 μg of the appropriate siRNA) were mixed (1:5 v/v)with i-Fect transfection reagent (Neuromics, Edina, MN). After recovery from the surgery (5–7 days), the animals received intrathecal injections (2 ug siRNA/10 ul/rat) of either a lipid encapsulated Raf-1-selective siRNA mixture (Smart pool siRNA, Dharmacon Inc; Chicago, IL, Cat # L-087699-00) (Raf-1 siRNA groups) or i-Fect encapsulated non-targeting dsRNA (Dharmacon, #D-001810-01-20) (control mismatch siRNA groups) or the transfection lipid alone, once daily, for 3 days, as described earlier (Gardell et al., 2002). Intrathecal injections of the siRNAs or the transfection agent alone did not cause any sign of behavioral toxicity. Western blots, using a Raf-1-selective antibody, indicated that intrathecal treatment with the Raf-1-selective siRNA mixture for 3 days significantly reduced Raf-1 protein levels in the dorsal root ganglion and in the dorsal horn of the spinal cord...
Louis Doré-Savard, Geneviève Roussy, Marc-André, Dansereau, Michael A Collingwood, Kim A Lennox, Scott D Rose, Nicolas Beaudet, Mark A Behlke and Philippe Sarret. Central Delivery of Dicer-substrate siRNA: A Direct Application for Pain Research. Molecular Therapy (2008); doi:10.1038/mt.2008.98
...In this study, we demonstrate the efficacy of 27-mer DsiRNA in reducing the expression of a specific G-protein coupled-receptor (GPCR) gene in rat spinal cord and DRG. Low doses of DsiRNA formulated in i-Fect, when administered by IT injection, induced a sustained reduction in the neurotensin receptor-2 (NTS2) GPCR mRNA and protein levels for 3–4 days. The reduction in NTS2 resulted in the expected behavioral changes in nociception. No apparent toxicity or nonspecific side effects were exhibited during the study period, and our results overall highlight the feasibility of using DsiRNA in pain research...
X.-W. Dong, S. Goregoaker, H. Engler, X. Zhou, L. Mark, J. Crona, R. Terry, J. Hunter and T. Priestley. Small interfering RNA-mediated selective knockdown of NaV1.8 tetrodotoxin-resistant sodium channel reverses mechanical allodynia in neuropathic rats. Neuroscience Volume 146, Issue 2, 11 May 2007, Pages 812-821
...For in vivo studies, siRNA stock solution (1 mg/ml) was prepared using siRNA suspension buffer (Qiagen). The solution was then heated to 90 °C for 1 min followed by 60 min incubation at 37 °C. Aliquots were stored at -20 °C. On the day of the injection, an aliquot of the siRNA was thawed on ice and then mixed with the transfection reagent, iFect (Neuromics Antibodies, Northfield, MN, USA), to yield a final concentration of 0.2 ug/ ul. A total of 100 ul of the siRNA solution or vehicle (20 ul of suspension buffer in 80 ul of iFect; 1:4 v/v) was locally delivered to lumbar DRGs through implanted epidural catheter by slow injection. The catheter was flushed with 15 ul sterile saline...
Luo MC, Zhang DQ, Ma SW, Huang YY, Shuster SJ, Porreca F, Lai J (2005). An efficient intrathecal delivery of small interfering RNA to the spinal cord and peripheral neurons. Molecular Pain 2005, 1:29.
Priti Kumar, Sang Kyung Lee, Premlata Shankar, N. Manjunath. A Single siRNA Cynthia C. Tsui, Stuart J. Shankland and Brian A. Pierchala. Glial Cell Line–Derived Neurotrophic Factor and Its Receptor Ret Is a Novel Ligand-Receptor Complex Critical for Survival Response during Podocyte Injury. doi: 10.1681/ASN.2005080835
...Ret small interference RNA (siRNA) knockdown was performed by using transient transfection of pooled functionally validated Ret siRNA (SMARTpool mouse RET siRNA; Dharmacon, Lafayette, CO). HSMP cells that were differentiated for 10 to 12 d were maintained at 10% FBS/RPMI as described above and transfected using the i-Fect siRNA transfection reagent (Neuromics, Edina, MN). For determination of the transfection efficiency, a Texas Red–labeled siRNA (siGLO RISCFree siRNA; Dharmacon) was co-transfected with Ret siRNA and visualized using fluorescence microscopy. For control siRNA samples, identical conditions were used with the substitution of siGLO-RISCFree siRNA for Ret siRNA. For determination of the efficiency of Ret knockdown, Western analysis for Ret was performed on WCL from cells 24 to 96 h after the transfection. Several concentrations of Ret siRNA (40, 60, and 100 nM) were tested to determine optimal knockdown conditions. For apoptosis assays, podocytes were exposed to UV-C or PA (40 g/ml) on days 3 to 4 after transfection of Ret siRNA or control siRNA (100 nM). Apoptosis was measured by counting podocytes with Hoechst-positive pyknotic nuclei 3 h after UV and 5 h after exposure to PA...
n-FectTM
Amy N. Sussman, Tong Sun, Ronald M. Krofft, and Raghu V. Durvasula. SPARC Accelerates Disease Progression in Experimental Crescentic Glomerulonephritis. Am. J. Pathol., May 2009; 174: 1827 - 1836.
...empty plasmid inserted into the NHe1/BamH1 sites of pcDNA3.1/Zeo () vector (Invitrogen, Carlsbad, CA) using N-Fect reagent (Neuromics, Edina, MN) according to manufacturers instructions. Transfected cells were selected in RPMI 1640 media supplemented with...
Hidetaka Sadanari, Junji Tanaka, Zhuan Li, Rie Yamada, Keiko Matsubara and Tsugiya Murayama.Proteasome inhibitor differentially regulates expression of the major immediate early genes of human cytomegalovirus in human central nervous system-derived cell lines. doi:10.1016/j.virusres.2009.01.010 .
King-Ho Cheung, Diana Shineman, Marioly Müller, César Cárdenas, Lijuan Mei1, Jun Yang, Taisuke Tomita, Takeshi Iwatsubo, Virginia M.-Y. Lee, and J. Kevin Foskett. Mechanism of Ca2+ Disruption in Alzheimer's Disease by Presenilin Regulation of InsP3 Receptor Channel Gating. doi:10.1016/j.neuron.2008.04.015
...pIRES2-EGFP empty vector by n-Fect reagent (Neuromics)...
Griffin SV, Hiromura K, Pippin J, Petermann AT, Blonski MJ, Krofft R, Takahashi S, Kulkarni AB, Shankland SJ (2004). Cyclin-Dependent Kinase 5 Is a Regulator of Podocyte Differentiation, Proliferation, and Morphology. Am J Pathol. 165:1175-1185.
Shih-Kuo Chen, Gladys Y.-P. Ko, and Stuart E. Dryer. Somatostatin Peptides Produce Multiple Effects on Gating Properties of Native Cone Photoreceptor cGMP-Gated Channels That Depend on Circadian Phase and Previous Illumination. J. Neurosci., Nov 2007; 27: 12168 - 12175 ; doi:10.1523/JNEUROSCI.3541-07.2007.
...Transfection was performed using pn-Fect from Neuromics (Edina, MN). Briefly, retinal cells were grown for 4 d in LD cycles, until 24 h before analysis. At that time, coverslips with attached retinal cells were transferred to 400 µl of serum-reduced culture medium (Optimem; Invitrogen, Carlsbad, CA) in 24-well plates supplemented with 100 µl of solution containing pn-Fect with PLC
-PH-GFP expression vector incorporated according to the manufacturer's instructions. In a series of pilot experiments, we determined that the maximum transfection efficiency occurred with a pn-Fect/DNA ratio of 1.4/1...